Development, Aging and Regeneration Program, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, USA.
Methods Mol Biol. 2020;2144:187-200. doi: 10.1007/978-1-0716-0592-9_17.
The cellular recycling process of autophagy is essential for survival, development, and homeostasis. Autophagy also plays an important role in aging and has been linked to longevity in many species, including the nematode C. elegans. Study of the physiological roles of autophagy during C. elegans aging requires methods for the spatiotemporal analysis of autophagy. Here we describe a method for assessing autophagic flux in multiple tissues of C. elegans by quantifying the pool of autophagic vesicles using fluorescently labelled Atg8/LGG-1 reporters upon autophagy inhibition using bafilomycin A (BafA). This methodology has revealed that autophagic activity varies in different cell types of C. elegans during aging.
自噬的细胞再循环过程对于生存、发育和内稳态至关重要。自噬在衰老过程中也起着重要作用，并与许多物种的长寿有关，包括线虫 C. elegans。研究自噬在 C. elegans 衰老过程中的生理作用需要时空分析自噬的方法。在这里，我们描述了一种通过在自噬抑制（使用 bafilomycin A，BafA）后使用荧光标记的 Atg8/LGG-1 报告器来定量自噬小体池，从而评估 C. elegans 多个组织中自噬通量的方法。该方法表明，自噬活性在 C. elegans 不同类型的细胞中随衰老而变化。
